Martin-Lewis Agar, Enriched

[所属分类:培养基配方] [发布时间:2021-5-17] [发布人:网站管理员2] [阅读次数:] [返回]

Martin-Lewis Agar, Enriched

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

Agar............................................................................................12.0g
Pancreatic digest of casein............................................................7.5g
Selected meat peptone ..................................................................7.5g
NaCl..............................................................................................5.0g
K 2 HPO 4 .........................................................................................4.0g
Cornstarch.....................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
Sarcina lutea suspension .........................................................20.0mL
Horse serum, inactivated .........................................................20.0mL
Supplement solution ................................................................10.0mL
PCAT inhibitor.........................................................................10.0mL
pH 7.2 ± 0.22 at 25°C
Source: The supplement solution (IsoVitaleX ® enrichment) is avail-
able from BD Diagnostic Systems. This enrichment may be replaced
by supplement VX from BD Diagnostic Systems.
Sarcina lutea Suspension:
Composition per 20.0mL:
Sarcina lutea FDA 1001.................................................10 6 –10 7 cells
Preparation of Sarcina lutea Suspension: Aseptically wash the
growth of 24-hr cultures of Sarcina lutea FDA 1001 cells from Thayer-
Martin plates with sterile soybean casein digest broth. Standardize the
suspension by adding additional sterile tryptic soy broth to yield 40%
light transmission at 530nm wavelength.
Soybean Casein Digest Broth:
Composition per liter:
Pancreatic digest of casein..........................................................17.0g
NaCl..............................................................................................5.0g
Papaic digest of soybean meal......................................................3.0g
K 2 HPO 4 .........................................................................................2.5g
Glucose.........................................................................................2.5g
pH 7.3 ± 0.2 at 25°C
Preparation of Soybean Casein Digest Broth: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–
121°C.
Supplement Solution:
Composition per liter:
Glucose .....................................................................................100.0g
L- Cysteine·HCl............................................................................25.9g
L- Glutamine.................................................................................10.0g
L- Cystine .......................................................................................1.1g
Adenine.........................................................................................1.0g
Nicotinamide adenine dinucleotide ............................................0.25g
Vitamin B 12 ...................................................................................0.1g
Thiamine pyrophosphate...............................................................0.1g
Guanine·HCl...............................................................................0.03g
Fe(NO 3 ) 3 ·6H 2 O...........................................................................0.02g
p-Aminobenzoic acid................................................................0.013g
Thiamine·HCl ............................................................................3.0mg
Preparation of Supplement Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter
sterilize.
PCAT Inhibitor:
Composition per 10.0mL:
Anisomycin.................................................................................0.02g
Colistin.......................................................................................7.5mg
Trimethoprim lactate..................................................................5.0mg
Penicillin G............................................................................25,000U
Preparation of PCAT Inhibitor: Add components to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Add components—except Sarcina lutea
suspension, horse serum, supplement solution, and PCAT inhibitor—to
distilled/deionized water and bring volume to 940.0mL. Gently heat
while stirring and bring to boiling. Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to 45°–50°C. Aseptically add 20.0mL of sterile Sarci-
na lutea suspension, 20.0mL of sterile horse serum, 10.0mL of
supplement solution, and 10.0mL of sterile PCAT inhibitor. Mix thor-
oughly. Pour into sterile Petri dishes.
Use: For the isolation and cultivation of pathogenic Neisseria, espe-
cially penicillinase-producing strains, from specimens containing
mixed flora of bacteria and fungi.